Objective To investigate the relationship between polycystic ovary syndrome (PCOS) susceptibility single nucleotide polymorphisms (SNP) and metabolic phenotypes in glucose and lipid metabolism and explore the pathophysiological mechanism of the susceptibility genes.Methods Three of PCOS susceptibility locus 2p16.3 (rs13405728 of LHCGR gene),2p21 (rs13429458,rs12478601 of THADA gene) and 9q33.3 (rs2479106,rs10818854 of DENNDIA gene) were selected and the metabolic phenotypes were compared between different genotypes of SNP in PCOS patients (using dominant model).Results Low-density lipoprotein cholesterol was significantly increased in CC genotype group than in TC + TT groups at rs12478601 of THADA gene [(2.5 ± 0.8),(2.4 ± 0.8) mmol/L; P =0.01].Serum insulin level of 2 hours after 75 g glucose intake was significantly higher in GG + AG groups than that of AA group at rs2479106 of DENND1A gene[(71 ±65),(64 ±50) mU/L;P =0.05],and the prevalence of type Ⅱ diabetes in first-degree relatives of patients were also increased [9.9％ (66/666),6.9％ (52/751) ; P ＜ 0.05].No association was found between metabolic phenotypes and genotypes of rs13429458,rs10818854,and rs13405728.Conclusions Genetic factors probably have effect on the metabolic characteristics of PCOS.THADA gene is related to lipid metabolism,while DENND1A gene may be involved in insulin metabolism in patients with PCOS.

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

This study aimed to investigate the synergistic effect of lidamycin (LDM) and rituximab on human B cell lymphoma Ramos cells. Cell proliferation was measured using MTS assay, cell apoptosis was analyzed by Annexin V-FITC/PI assay, the expression of apoptosis related proteins was analyzed by Western blotting, and the in vivo lymphoma inhibition was verified using BALB/c mice inoculated via tail vein using Ramos cells which stably expressed pEGFP-N1 plasmid. The results showed that, after the pretreatment with rituximab for 48 h, rituximab and LDM showed significantly synergistic effects on cell proliferation. Cells in combined treatment group had a higher apoptosis rate than that in LDM treatment group. Compared with the LDM treatment group, the expression of apoptosis-related proteins such as Cleaved caspase-3, Cleaved caspase-7, Cleaved caspase-9 and Cleaved PARP in combined treatment groups increased, and expression of cIAP-2 and Bcl-2 decreased. The result of in vivo experiment showed that, in the combined treatment group, the survival time of BALB/c mice was significantly longer than the mice in control group and LDM treatment group, and the degree of tumor accumulation and metastasis to lymph nodes and spleen was lower.
Aminoglycosides ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 7 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Synergism ; Enediynes ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Lymphoma, B-Cell ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rituximab ; pharmacology

Objective To explore the clinicopathological characteristics of hereditary ovarian cancer syndrome(HOCS). Methods From Jan. 2000 to Jan. 2007, among 580 cases of primary ovarian cancer, 42 cases(herediatary group),who had a positive family history of ovarian cancer and met the diagnostic criteria of HOCS, were analyzed retrospectively. One hundred cases without a family history of ovarian cancer were enrolled randomizely as control group (sporadic group). Results The incidence of HOCS was 7.2% (42/580). Forty-two cases associated tumors affected at least 2 successive generations in 31 families and affected 1 generation in 8 families. Eighty-seven percent (27/31)was from maternal lineage, while 13% (4/31)from paternal lineage. Earlier age of onset was significantly difference between two groups[(49±10) years vs. (55±10) years, P<0.05]. There were 90% belong to serous adenocarcinoma in the herediatary group, while 84% in the sporadic group. There was statistical difference in the proportion of mucinous adenocarcinoma (0 vs. 11%, P<0.05). The most common clinical manifestations were abdominal distention and anorexia (64% vs. 70%, P>0.05), International Federational of Gynecology Obstetrics(FIGO)stage Ⅲ (62% vs. 63%, P>0.05) between two groups. Fourteen cases (33%,14/42) were previously untreated in the herediatary group, while 40 cases (40%, 40/100) in the sporadic group. There were 15 cases (36%, 15/42) underwent secondary surgery and 15 cases (36%, 15/42) underwent third surgery or more in berediatary group, while 50 cases (50%, 50/100) and 27 cases(27%, 27/100) in the sporadic group. The mean number of ehemotberapy cycles received in two groups was 13.3 and 11.8 (P>0.05). The 3-year and 5-year survival rate in herediatary group were 73.6% and 54.9% respectively, compared with 47.4% and 21.2% (P<0.05) in sporadic group. Conclusion Hereditary ovarian cancer mostly from maternal lineage are featuring in early age of onset, serous adenocarcinoma, advanced stage (stage Ⅲ), and better prognosis after the comprehensive treated by cytoreductive surgery plus with chemotherapy.

Objective To investigate cerebral MRI characteristics of patients with eclampsia. Methods A retrospective study was conducted on 15 cases of eclampsia and items reviewed cover all data concerning clinical features,cerebral MRI findings and results of follow-up survey.Results All of these patients had clinical symptoms of.blurred vision,headache,seizure,hypertension,proteinuria and edema of lower extremity.As for the characteristics of imaging,13 cases had only abnormal symmetric signals in parieto-occipital lobes,frontal lobe and basal ganglia were involved in 2 cases,and temporal lobe was involved in 1 case.The signals of lesions in DWI were isointense or hypointense,however they were hyperintense in ADC map.Two cases had hyperintense signals in DWI.All the patients recovered well,and all brain lesions disappeared during follow up.Conclusion The most important imaging of eclampsia is vasogenic edema with a good prognosis.

OBJECTIVETo investigate the effect of suppression of CCL5 ligand gene on the proliferation of human breast cancer cells.

METHODSA lentiviral vector carrying a short interfering RNA (siRNA) targeting CCL5 was transfected into human breast cancer cell line MCF-7 and MDA-MB-231 cells. The expression of CCL5 mRNA in the cells was detected by real-time PCR. The proliferation of MCF-7 and MDA-MB-231 cells was assessed by MTT assay and FACS assay, and the colony formation ability of both cell lines were measured, respectively.

RESULTSReal time PCR showed a good knockdown effect of CCL5 in both cell-lines. Colony-forming assay showed that the ability of colony formation of MCF-7/CCL5-siRNA and MDA-MB-231/CCL5-siRNA was decreased markedly. The colony number of MCF-7/CCL5-siRNA group was (0.34 ± 0.08), significantly lower than 0.81 ± 0.12 in the MCF-7/CCL5-N group and 0.92 ± 0.12 in the MCF-7 group (P < 0.05). The colony number of MDA-MB-231/CCL5-siRNA group was 0.33 ± 0.10, significantly lower than 0.97 ± 0.09 in the MDA-MB-231/CCL5-N group and 1.04 ± 0.07 in the MDA-MB-231 group (P < 0.05). However, MTT assay revealed that the proliferation of MCF-7/CCL5-siRNA cells was not significantly different from that of MCF-7/CCL5-N or MCF-7 cells, respectively (P > 0.05), and the same result was found in MDA-MB-231 cells. FACS assay showed that the proliferation index (PI) of groups MCF-7/CCL5-siRNA, MCF-7/CCL5-N and MCF-7 were 0.48 ± 0.03, 0.43 ± 0.01 and 0.45 ± 0.02. The PI of groups MDA-MB-231/CCL5-siRNA, MDA-MB-231/CCL5-N and MDA-MB-231 cells were 0.48 ± 0.02, 0.44 ± 0.05 and 0.47 ± 0.02. There was no statistical difference among them (P > 0.05).

CONCLUSIONThe down-regulation of CCL5 gene in human breast cancer cells may significantly suppress their colony formation ability, rather than affecting their population doubling time to some extent.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Chemokine CCL5 ; genetics ; metabolism ; physiology ; Down-Regulation ; Female ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo study the mechanism of treatment of post liposuction seroma with Staphylococcal enterotoxin C injection.

METHODS64 cases with post liposuction seroma were treated with Staphylococcal enterotoxin C injection or routine procedures. The exudate of those patients was collected to analyze the ratio, pH value, cell species and numbers, and the value of TP, ALP, LDH, AST, ALT, gamma-GT, ADA, ApoB, TC.

RESULTSThe ratio, numbers of lymphocyte and mesothelial cells and TP, LDH, ADA, TC value in exudate in Staphylococcal enterotoxin C group were significantly higher than those in control group.

CONCLUSIONSThe effect of Staphylococcal enterotoxin C injection on the exudate of seroma may be related to the non-inflammation reaction.

Enterotoxins ; therapeutic use ; Female ; Humans ; Lipectomy ; adverse effects ; Postoperative Complications ; drug therapy ; metabolism ; Seroma ; drug therapy ; etiology ; metabolism

Endoplasmic reticulum stress (ERS) is an adaptive process in response to circumstantial changes, but excessive and/or prolonged ERS can induce cell apoptosis. C/EBP homologous protein (CHOP) is a very important marker participating in ERS-associated cell apoptosis, while the role of the myocyte apoptosis induced by CHOP remains unclear in the development of hypertrophy. The present study aimed to investigate the effect of CHOP-mediated ERS-associated apoptosis on myocardial hypertrophy induced by abdominal aortic constriction in rats. Healthy male Wistar rats were randomly divided into model group (n=45) and control group (n=40). The rats in model group received abdominal aortic constriction. Hemodynamic changes, whole heart weight/body weight (HW/BW) and left ventricular weight/body weight (LVW/BW) were measured on 1 d, 3 d, 7 d, 14 d and 28 d after surgery, respectively. The mRNA expression of glucose-regulated protein 78 (GRP78), calreticulin (CRT) and CHOP, which are important markers of ERS, were detected by RT-PCR, and Western blot was used to assess the protein level of GRP78, CRT, CHOP, and apoptosis-associated proteins, Bax and Bcl-2. The results obtained were as follows. Compared with control group, the blood pressure, LVW/BW, and HW/BW of rats in model group increased significantly and cardiac function enhanced compensatively on 7 d after surgery, and increased progressively during the experiment. As early as 1 d after surgery, the mRNA level of CRT in model group increased by 136% (P< 0.01) compared with control, while the protein expression increased by 69.2% on 7 d after surgery (P<0.01). Both mRNA and protein expression of GRP78 increased by 20% and 186% (P<0.01) respectively on 7 d after surgery, and the expression sustained high level during the experiment afterwards. Correlation analysis indicated a positive correlation between +dp/dt(max) and CRT protein expression (r=0.780, P<0.01) as well as GRP78 protein expression (r=0.694, P<0.01). Prolonged ERS triggered myocyte apoptosis, as both the mRNA and protein level of CHOP in model group increased by 22.2% (P<0.01) and 76.0% (P<0.01) respectively compared with control on 7 d after hypertrophy (14 d after surgery), and meanwhile, the protein expression of pro-apoptotic Bax increased by 41.1% (P<0.01) and anti-apoptotic Bcl-2 protein expression decreased by 25.5% (P<0.01). Correlation analysis indicated a positive correlation between CHOP and Bax expression (r=0.654, P<0.01), and a negative correlation between CHOP and Bcl-2 expression (r=-0.671, P<0.01). These results suggest that abdominal aortic constriction induces a significant up-regulation in ER molecular chaperones at early stage of post-surgery, indicating that ERS response is activated in the rat heart; while prolonged ERS could lead to myocyte apoptosis, and CHOP-mediated ERS-associated apoptosis may contribute to myocardial hypertrophy. We speculate that cell apoptosis may take part in the regulation of myocardial hypertrophy and heart failure, and determine the progression of decompensated hypertrophy.
Animals ; Aorta ; physiopathology ; Apoptosis ; Calreticulin ; metabolism ; Constriction ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Hypertrophy ; pathology ; Male ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Transcription Factor CHOP ; metabolism ; Up-Regulation ; bcl-2-Associated X Protein ; metabolism

OBJECTIVEHepatic stellate cells (HSC) in hepatocellular carcinoma (HCC) transdifferentiate into extracellular matrix-producing myofibroblasts. Activated HSC can promote invasion and metastasis of HCC. To understand the differences of HSC in normal liver and HCC, we compared the gene expression patterns in HCC cell induction-activated and culture-activated rat HSC.

METHODSHSC were isolated by density centrifugation and exposed to conditioned medium from rat HCC cell line C5F. Expression of 22 012 genes in quiescent HSC, culture-activated HSC and HCC induction-activated HSC was analyzed by cDNA microarray and confirmed by real-time RT-PCR and Western blot.

RESULTS1672 genes were differentially expressed in culture-activated HSC, including proinflammatory factors, cell adhesion molecules, cell surface receptors, signaling transduction molecules and immune factors. 711 genes were differentially expressed in HCC induction-activated HSC. Some of them were identical to those in culture-activated HSC. HCC Induction-activated HSC showed specific gene expression patterns, including Raf1, Rac2, Adam17, Wnt6, MMP-9 and TNF, suggesting that HCC cells can specifically induce HSC activation.

CONCLUSIONThe gene expression patterns in HCC induction-activated HSC are different from those in culture-activated HSC. HCC induction-activated HSC may play a major role in the invasion and metastasis of HCC. In vivo activation should be considered as the standard for the study of HSC biology. HCC induction-activated HSC should be considered as the standard for HSC biology studies.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Hepatic Stellate Cells ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Inbred F344

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism